摘要
目的:追踪附片液制备过程中14个生物碱的传递规律。方法:采用Phenomenonex Gemini C18色谱柱(150 mm×4. 6 mm,5μm),流动相为0. 1%甲酸水溶液(A)-乙腈(B),质谱采用ESI+多反应监测(multiple reaction monitoring,MRM)模式扫描;建立HPLC-MS/MS方法同时测定附片液中乌头碱、新乌头碱、次乌头碱、印乌头碱、苯甲酰乌头原碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱、乌头原碱、尼奥林、附子灵、宋果灵、塔拉乌头胺、去甲乌药碱、去甲猪毛菜碱14个生物碱含量的检测方法;结合聚类分析、主成分分析追踪各种生物碱在附片液制备过程中8个环节的传递规律。结果:14个生物碱在各自浓度范围内线性关系良好(R^2> 0. 990 0),定量限为2. 27～18. 27 ng·mL^(-1),平均回收率94. 81%～101. 55%。分析表明,煎煮过程中各种生物碱大量溶出,剧毒的乌头碱、新乌头碱与印乌头碱全部水解完毕,次乌头碱含量亦水解降低了63. 5%,大量单酯型生物碱转化生成,含量显著升高;浓缩环节各种生物碱均有不同程度损失,次乌头碱含量进一步降低至饮片总量的15. 4%,去甲猪毛菜碱损失超过63. 9%;通过2次醇沉精制环节,大量去除提取液中的淀粉、多糖等物质,而对各种生物碱含量影响不大。结论:附片液在制备过程中,毒性成分基本水解完毕,保证了制剂的安全性,但浓缩环节成分损失较大,应控制温度,改进工艺。该研究为附片液生产工艺的优化及品质提升提供了科学依据。
Objective:To investigate the transfer rule of fourteen alkaoids in the preparation process of Fupian liquid.Methods:Phenomenonex Gemini C18 column (150mm×4.6mm,5μm)was used and 0.1% formic acid aqueous solution(A)-acetonitrile (B)was selected as mobile phase.The mass spectrum was scanned by ESI^+ multiple reaction monitoring (MRM)mode.An HPLC-MS/MS method was established for the simultaneous determination of aconitine,mesaconitine,hypaconitine,indaconitine,benzoylaconine,benzoylmesaconine, benzoylhypaconitine,aconine,fuziline,neoline,talatisamine,songorine,higenamine and salsoline in the preparation process of Fupian liquid.Combined with cluster analysis and principal component analysis,the transfer rule of various alkaoids was tracked in the eight steps of aconite oral liquid preparation.Results :Methodological validation results showed that the linearity of the calibration curves of the 14 compounds was good (R^2>0.990 0).The limits of quantification were 2.27-18.27ng·mL^-1 ,and the average recoveries were 94.81%-101.55%.The analysis results illustrated that a large number of alkaloids dissolved in the decocting step.Toxic ingredients,such as aconitine, mesaconine and dexyaconitine,were completely hydrolyzed.And the content of hypaconitine was also reduced to 63.5% by hydrolysis.Due to the hydrolysis of diester alkaloids,a large number of monoester alkaloids were produced and their contents were obviously increased.In the concentration step,the 14 alkaloids had different degrees of loss.Among them,the content of hypaconitine was further reduced to 15.4% and the loss of salsoline was more than 63.9%.After two ethanol precipitation steps,a number of starch,polysaccharides and other substances were removed from the extract,but there was little effect on the contents of alkaoids.Conclusion:In the preparation process of Fupian liquid,toxic ingredients are all hydrolyzed,which ensures the drug safety. However,in the concentration step,the loss of active ingredients is impressive,the temperature should be controlle
作者
贺亚男
熊茜
林俊芝
潘媛
李燕
韩丽
杨明
许润春
张定堃
HE Ya-nan;XIONG Xi;LIN Jun-zhi;PAN Yuan;LI Yan;HAN Li;YANG Ming;XU Run-chun;ZHANG Ding-kun(Key Laboratory Breeding Base of Systematic Research and Utilization on Chinese Material Medical Resources Co-founded By Sichuan Province and Ministry of Science and Technology,Pharmacy College, Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Teaching Hospital of Chengdu University of Traditional Chinese Medicine,Central Laboratory,Chengdu 610075,China;Testing and Analysis Center,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;State Key Laboratory of Innovation Drug and Efficient Energy-Saving Pharmaceutical Equipment, Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2018年第24期2954-2961,共8页
Chinese Journal of New Drugs
基金
四川省中医药管理局科研专项(2018QN008)
成都中医药大学科技发展基金资助项目(ZRQN1515)
